"
Team:DTU-Denmark/Notebook/5 August 2013
From 2013.igem.org
(→Results) |
(→Procedure) |
||
| Line 27: | Line 27: | ||
Samples 1-5 and 8 were top10 transformant colonies. Colonies 1-5 were small (1mm wide) more condense and more difficult to collect; this may be due to the proteins expressed on the membrane that make it thicker. Colony 8 was wider (3mm wide) and more sticky. | Samples 1-5 and 8 were top10 transformant colonies. Colonies 1-5 were small (1mm wide) more condense and more difficult to collect; this may be due to the proteins expressed on the membrane that make it thicker. Colony 8 was wider (3mm wide) and more sticky. | ||
Samples 6 and 7 are normal transformant colonies. | Samples 6 and 7 are normal transformant colonies. | ||
| + | |||
| + | ===Colony PCR === | ||
| + | |||
| + | To check HAO transformants from USER cloning on the 01.08. | ||
| + | |||
| + | ===Nanodrop=== | ||
| + | |||
| + | Measured Midiprep from 03.08.. No DNA was detectable in the samples. Redo purification. | ||
| + | |||
| + | ===PCR=== | ||
| + | |||
| + | PCR for araBAD with new RV primer, SPL (synthetic promoter library) in pZA21 and reference promoter in pZA21. | ||
| + | |||
| + | samples: | ||
| + | * ara1, ara 2 - primers 12a + 12bn, template K808000, 57C, 1:00 | ||
| + | * spl1, spl2 - primers 52a + 52b1, template RFP in pZA21, 54C, 2:30 | ||
| + | * ref1, ref2 - primers 52a + 52b2, template RFP in pZA21, 54C, 2:30 | ||
==Results== | ==Results== | ||
Revision as of 15:27, 5 August 2013
5 August 2013
Contents |
lab 208
Main purpose
- Colony PCR for HAO
- USER reaction
Who was in the lab
Kristian, Julia, Natalia, Henrike
Procedure
Gels
Run 1% agarose gel on colony PCR products. The samples are named:
- HAO 1
- HAO 2
- HAO 3
- HAO 4
- HAO 5
- HAO 6
- HAO 7
- HAO 8
Samples 1-5 and 8 were top10 transformant colonies. Colonies 1-5 were small (1mm wide) more condense and more difficult to collect; this may be due to the proteins expressed on the membrane that make it thicker. Colony 8 was wider (3mm wide) and more sticky. Samples 6 and 7 are normal transformant colonies.
Colony PCR
To check HAO transformants from USER cloning on the 01.08.
Nanodrop
Measured Midiprep from 03.08.. No DNA was detectable in the samples. Redo purification.
PCR
PCR for araBAD with new RV primer, SPL (synthetic promoter library) in pZA21 and reference promoter in pZA21.
samples:
- ara1, ara 2 - primers 12a + 12bn, template K808000, 57C, 1:00
- spl1, spl2 - primers 52a + 52b1, template RFP in pZA21, 54C, 2:30
- ref1, ref2 - primers 52a + 52b2, template RFP in pZA21, 54C, 2:30
Results
1% agarose gel
- 1kb ladder
- HAO 1 (from top 10 transformants)
- HAO 2
- HAO 3
- HAO 4
- HAO 5
- HAO 6
- HAO 7
- HAO 8
- 1 kb ladder
1% agarose gel on PCR products
Kristian loaded it :)
Conclusion
Navigate to the Previous or the Next Entry
